Establishment of serodiagnostic method and surveylance system for hemorrhagic fever with renal syndrome virus infection in experimental animals

实验动物肾综合征出血热病毒感染血清学诊断方法及调查体系的建立

• 批准号: 06558112
• 负责人: ARIKAWA Jiro
• 金额: $ 11.84万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06558112/
• 关键词: HFRS; Hantavirus; Baculovirus; Serological diagnosis; ELISA; Westem blotting; Experimental animals; Zoonosis

1. Recombinant baculovirus expressing hantavirus nucleocapsid protein (Bac.HTN-NP) was applied for the antigen of ELISA.Specificity of the ELISA was the same to that of the IFA which uses infected Vero E6 cells as antigen. However, the ELISA had a sensitivity problem because of the relatively high background coloring.2. cDNA which encoded hantavirus nucleocapsid protein was expressed by using E.coli system and established capture ELISA system. This ELISA was shown to have same specificity and sensitivity to that of the ordinally IFA,and therefore, considered as a useful screening method.3. Bac.HTN-NP was successfully applied for the antigen for Western blotting (WB). The WB method readily distinguished specific reaction from nonspecific reaction from the WB pattern. Therefore, the WB was considered as an effective method for the serologic confirmation of hantavirus infection.

1.用表达汉坦病毒核衣壳蛋白的重组杆状病毒(Bac.HTN-NP)作为ELISA的抗原，其特异性与以感染的Vero E6细胞为抗原的IFA相同。但是，由于背景显色相对较高，该方法存在敏感性问题。利用大肠杆菌系统表达了汉坦病毒核衣壳蛋白基因，并建立了捕获酶联免疫吸附试验体系。该方法具有与常规IFA相同的特异性和敏感性，是一种有效的筛查方法。成功地将Bac.HTN-NP作为免疫印迹(WB)的抗原。WB方法很容易从WB模式中区分特定反应和非特定反应。因此，WB被认为是一种有效的汉坦病毒感染的血清学确认方法。

项目成果

Jiro Arikawa: "Epizootiological studies of hantavirus infection among urban rats in Hokkaido,Japan:Evidences for the persistent infection from the sero-epizootiological surveys and antigenic characterizations of hantavirus isolates." J.Vet.Med.Sci.56. 27-

Jiro Arikawa：“日本北海道城市老鼠汉坦病毒感染的流行病学研究：来自血清流行病学调查和汉坦病毒分离株抗原特征的持续感染证据。”

Yoo, YC.: "Effect of MDP-Lys (L18), a derivatives of MDP, on enhancing host resistance against Hantaan virus infection in newborn mice." Vaccine. 13. 1300-1305 (1995)

Yoo, YC.：“MDP-Lys (L18)（MDP 的衍生物）对增强新生小鼠宿主对汉坦病毒感染的抵抗力的作用。”

Yoshimatsu, K., Arikawa, J., Tamura, M., Yoshida, R., Lundkvist, A., Niklasson, B., Kariwa, H., Azuma, I.: "Characterization of the nucleocapsid protein of hantaan virus strain 76-118 using monoclonal antibodies." J.Gen.Virol.77. 695-704 (1996)

Yoshimatsu, K.、Arikawa, J.、Tamura, M.、Yoshida, R.、Lundkvist, A.、Niklasson, B.、Kariwa, H.、Azuma, I.：“汉坦病毒株核衣壳蛋白的表征

Yoshimatsu,K.: "Characterization of the nucleoapsid protein of Hantaan virus strain 76-118 using monoclonal antibodies" Journal of General Virology. 77. 695-704 (1996)

Yoshimatsu,K.：“使用单克隆抗体表征汉滩病毒株 76-118 的核壳蛋白”《普通病毒学杂志》。

Yoshimatsu, K.: "Western blotting using recombinant Hantaan virus nucleocapsid protein expressed in silkworm as a serological confirmation of hantavirus infection in human sera." Journal of Veterinary Medical Science. 58. 71-74 (1996)

Yoshimatsu, K.：“使用在家蚕中表达的重组汉坦病毒核衣壳蛋白进行蛋白质印迹，作为人类血清中汉坦病毒感染的血清学确认。”